Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia.
Int J Food Microbiol. 2014 Feb 17;172:92-101. doi: 10.1016/j.ijfoodmicro.2013.11.034. Epub 2013 Dec 7.
In this study, a Salmonella Typhimurium lytic bacteriophage, Φ st1, which was isolated from chicken faecal material, was evaluated as a candidate for biocontrol of Salmonella in chickens. The morphology of Φ st1 showed strong resemblance to members of the Siphoviridae family. Φ st1 was observed to be a DNA phage with an estimated genome size of 121 kbp. It was found to be able to infect S. Typhimurium and S. Hadar, with a stronger lytic activity against the former. Subsequent characterisation of Φ st1 against S. Typhimurium showed that Φ st1 has a latent period of 40 min with an average burst size of 22 particles per infective centre. Approximately 86.1% of the phage adsorbed to the host cells within the initial 5 min of infection. At the optimum multiplicity of infection (MOI) (0.1), the highest reduction rate of S. Typhimurium (6.6 log₁₀ CFU/ml) and increment in phage titre (3.8 log₁₀ PFU/ml) was observed. Φ st1 produced adsorption rates of 88.4-92.2% at pH7-9 and demonstrated the highest bacteria reduction (6.6 log₁₀ CFU/ml) at pH9. Φ st1 also showed an insignificant different (P>0.05) reduction rate of host cells at 37 °C (6.4 log₁₀ CFU/ml) and 42 °C (6.0 log₁₀ CFU/ml). The in vivo study using Φ st1 showed that intracloacal inoculation of ~10¹² PFU/ml of the phage in the chickens challenged with ~10¹⁰ CFU/ml of S. Typhimurium was able to reduce (P<0.05) the S. Typhimurium more rapidly than the untreated group. The Salmonella count reduced to 2.9 log₁₀ CFU/ml within 6h of post-challenge and S. Typhimurium was not detected at and after 24h of post-challenge. Reduction of Salmonella count in visceral organs was also observed at 6h post-challenge. Approximately 1.6 log₁₀ FU/ml Φ st1 was found to persist in the caecal wall of the chicks at 72 h of post-challenge. The present study indicated that Φ st1 may serve as a potential biocontrol agent to reduce the Salmonella count in caecal content of chickens.
在这项研究中,从鸡粪便中分离出的一株肠炎沙门氏菌裂解噬菌体 Φst1 被评估为控制鸡中沙门氏菌的候选生物防治剂。 Φst1 的形态与肌尾噬菌体科的成员非常相似。 Φst1 被观察为一种 DNA 噬菌体,估计基因组大小为 121 kbp。它能够感染肠炎沙门氏菌和肠炎哈达,对前者具有更强的裂解活性。随后对 Φst1 进行的肠炎沙门氏菌特性分析表明, Φst1 的潜伏期为 40 分钟,平均每个感染中心有 22 个粒子爆发。大约 86.1%的噬菌体在感染的最初 5 分钟内吸附到宿主细胞上。在最佳感染复数 (MOI) (0.1) 下,观察到肠炎沙门氏菌的最高减少率 (6.6 log₁₀ CFU/ml) 和噬菌体滴度的最高增加 (3.8 log₁₀ PFU/ml)。 Φst1 在 pH7-9 时产生 88.4-92.2%的吸附率,并在 pH9 时表现出最高的细菌减少 (6.6 log₁₀ CFU/ml)。 Φst1 在 37°C (6.4 log₁₀ CFU/ml) 和 42°C (6.0 log₁₀ CFU/ml) 下对宿主细胞的减少率也没有显著差异 (P>0.05)。使用 Φst1 的体内研究表明,在受到约 10¹⁰ CFU/ml 肠炎沙门氏菌挑战的鸡中,经腔内接种约 10¹² PFU/ml 的噬菌体,可使肠炎沙门氏菌的减少速度快于未处理组 (P<0.05)。肠炎沙门氏菌的数量在攻毒后 6 小时内减少到 2.9 log₁₀ CFU/ml,攻毒后 24 小时及之后均未检测到肠炎沙门氏菌。攻毒后 6 小时也观察到内脏器官中肠炎沙门氏菌数量的减少。在攻毒后 72 小时,鸡盲肠壁中约有 1.6 log₁₀ FU/ml 的 Φst1 持续存在。本研究表明, Φst1 可作为一种潜在的生物防治剂,减少鸡盲肠内容物中肠炎沙门氏菌的数量。